Manual de procedimentos básicos em microbiologia



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PACIENTES NEUTROPÊNICOS E IMUNOSSUPRIMIDOS


Além dos agentes relatados como causa de pneumonia em crianças e adultos, devem ser valorizados achados clinicamente compatíveis de:



  • Bactérias: Streptococcus viridans, Corynebacterium jeikeium, Bacillus spp., Legionella spp., Mycobacterium spp., Nocardia spp., Rodococcus spp.

  • Fungos: Aspergillus spp., Fusarium spp., Candida spp., P. carinii, Cryptococcus neoformans

  • Protozoários: Toxoplasma gondii, Strongyloides stercoralis



processamento microbiológico de amostras

Meios recomendados para a cultura das amostras do trato respiratório:



  • Ágar sangue

  • Ágar Mac conkey

  • Ágar chocolate

  • Ágar sangue suplementado para anaeróbios, para amostras clínicas para as quais recomenda-se fazer o isolamento de anaeróbios.

  • Quando é indicada cultura para Legionella spp., fungos, micobactérias, Chlamydia e vírus, acrescentam-se os meios necessários a estas rotinas específicas. A pesquisa por imunofluorescência com anticorpos monoclonais, e os métodos moleculares são mais recomendados para a detecção desses microrganismos.

A semeadura da amostra e interpretação do número de colônias no caso da utilização de técnicas quantitativas, poderá ser feita de uma das formas abaixo:




    • Após homogeneização da amostra, semear 10 µl, diretamente nas placas, utilizando-se alças calibradas descartáveis ou pipeta com ponteiras estéreis.




Nº de colônias na placa após Incubação “overnight”

Interpretação em UFC/ml

< 10

< 103

10 a 100

103 a 104

100 a 1000

104 a 105


Observação: Para o Lavado broncoalveolar considerar a diluição pelo volume injetado e multiplicar por 100. P.ex., se a leitura for 50 colonias, multiplicar por 100 (volume da alça) e multiplicar por 100 (diluição da coleta). Resultado final = 5x10x 10 2x10 2 = 5 x 105 UFC/ml


    • Usar diluições de:

  • 1/10: 10 µl com alça calibrada semeada no Ágar Chocolate

  • 1/100: 1 ml do material diluído em 9 ml de solução salina e semeando-se 10 µL desta solução com alça calibrada na placa de Ágar Chocolate

  • 1/1000: usar a solução anterior e semear 1 µL com alça calibrada na placa de Ágar Mac Conkey.

Para expressão do resultado em ml o número de colônias obtido deverá ser corrigido pelo fator da diluição e correlacionado com a quantidade de amostra semeada.




  • prepara-se uma diluição de 1:20 (0,5 ml de fluido em 9,5mL de solução salina estéril). Deste caldo semeia-se 50 µl em cada um dos meios selecionados.




Nº de colônias na placa após incubação “overnight”

Interpretação em UFC/ml


2-24

103

25-249

104

≥ 250

105


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